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血管活性腸肽(VIP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Vasoactive Intestinal Peptide (VIP)

PHM27; Intestinal peptide PHV-42; Peptide histidine valine 42; Peptide histidine methioninamide 27

  • 血管活性腸肽(VIP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 血管活性腸肽(VIP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 血管活性腸肽(VIP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 實(shí)驗(yàn)結(jié)果圖
  • CEA380Bo.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)血管活性腸肽(VIP),經(jīng)檢測(cè)與其它相似物質(zhì)無(wú)明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來(lái)源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)的其它物質(zhì)有交叉反應(yīng)。

回收率

分別于定值血清及血漿樣本中加入一定量的血管活性腸肽(VIP)(加標(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。

樣本 回收率范圍(%) 平均回收率(%)
serum(n=5) 80-99 90
EDTA plasma(n=5) 86-95 91
heparin plasma(n=5) 82-104 91

精密度

精密度用樣品測(cè)定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)算不同濃度樣本的平均值及SD值。
批內(nèi)差: CV<10%
批間差: CV<12%

線性

在定值血清及血漿樣本內(nèi)加入適量的血管活性腸肽(VIP),并倍比稀釋成1:2,1:4,1:8,1:16的待測(cè)樣本,線性范圍即為稀釋后樣本中血管活性腸肽(VIP)含量的測(cè)定值與理論值的比率。

樣本 1:2 1:4 1:8 1:16
serum(n=5) 78-97% 96-105% 79-95% 78-94%
EDTA plasma(n=5) 78-97% 90-101% 81-94% 79-94%
heparin plasma(n=5) 78-94% 78-95% 79-92% 96-103%

穩(wěn)定性

經(jīng)測(cè)定,試劑盒在有效期內(nèi)按推薦溫度保存,其活性降低率小于5%。
為減小外部因素對(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來(lái)進(jìn)行操作可減少人為誤差。

實(shí)驗(yàn)流程

1. 實(shí)驗(yàn)前標(biāo)準(zhǔn)品、試劑及樣本的準(zhǔn)備;
2. 加樣(標(biāo)準(zhǔn)品及樣本)50µL,
    加入50µL檢測(cè)液A(臨用前配制);
    37°C溫育1小時(shí)。
3. 洗板3次;
4. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
5. 洗板5次;
6. 加TMB底物90µL,37°C孵育10-20分鐘;
7. 加終止液50µL,立即450nm讀數(shù)。

實(shí)驗(yàn)原理

本試劑盒應(yīng)用競(jìng)爭(zhēng)抑制酶聯(lián)免疫分析法測(cè)定標(biāo)本中待測(cè)物質(zhì)水平。將血管活性腸肽(VIP)單克隆抗體包被微孔板,制成固相載 體,往包被抗體的微孔中同時(shí)加入生物素標(biāo)記的抗原和待測(cè)抗原(標(biāo)準(zhǔn)品或樣本),待測(cè)抗原與生物素標(biāo)記抗原對(duì)特異性抗體進(jìn)行競(jìng)爭(zhēng)結(jié)合。溫育后經(jīng)洗滌去掉未結(jié)合物,然后加入HRP標(biāo)記的親和素,經(jīng)過溫育和徹底洗滌后加入底物TMB顯色。TMB在過氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成最終的黃色。待測(cè)標(biāo)本濃度越高,標(biāo)記抗原和抗體的結(jié)合就越受到抑制,顯色愈淺。顯色的深淺與酶量呈正相關(guān),而與樣品中待測(cè)物質(zhì)含量呈負(fù)相關(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。

相關(guān)產(chǎn)品

編號(hào) 適用物種:Bos taurus; Bovine (Cattle,牛) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
RPA380Bo01 血管活性腸肽(VIP)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
PAA380Bo01 血管活性腸肽(VIP)多克隆抗體 WB; IHC; ICC; IP.
CEA380Bo 血管活性腸肽(VIP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA380Bo 血管活性腸肽(VIP)等多因子檢測(cè)試劑盒(流式熒光發(fā)光法) FLIA Kit for Antigen Detection.

參考文獻(xiàn)

雜志 參考文獻(xiàn)
Clinical Genetics VIP gene variants related to idiopathic pulmonary arterial hypertension in Chinese population [Wiley: source]
Human immunology Low levels of vasoactive intestinal peptide are associated with Chagas disease cardiomyopathy [Pubmed: 23800435]
Headache: The Journal of Head and Face Pain CGRP and VIP Levels as Predictors of Efficacy of Onabotulinumtoxin Type A in Chronic Migraine [Onlinelibrary: head.12372]
World J Gastroenterol Roles of sphincter of Oddi motility and serum vasoactive intestinal peptide, gastrin and cholecystokinin octapeptide [NCBI: PMC4000510]
Cephalalgia? Increased VIP levels in peripheral blood outside migraine attacks as a potential biomarker of cranial parasympathetic activation in chronic migraine [Pubmed:24847167]
J Huazhong Univ Sci Technolog Med Sci. Clinical study of electro-acupuncture treatment with different intensities for functional constipation patients [Pubmed:25318892]
PLOS ONE Aqueous Fraction of Beta vulgaris Ameliorates Hyperglycemia in Diabetic Mice due to Enhanced Glucose Stimulated Insulin Secretion, Mediated by Acetylcholine and GLP-1, and Elevated Glucose Uptake via Increased Membrane Bound GLUT4 Transporters [Pubmed:25647228]
PLOS ONE Aqueous Fraction of [PubMed: 25647228]
Pharmacol Res Oral curcumin has anti-arthritic efficacy through somatostatin generation [PubMed: 25836921]
World J Gastroenterol Effects of sphincter of Oddi motility on the formation of cholesterol gallstones [Pubmed:27350732]
Annals of Allergy, Asthma and Immunology Effect of acupuncture on house dust mite specific IgE, substance P, and symptoms in persistent allergic rhinitis [Pubmed:27156748]
Cephalalgia. Relationship between serum levels of VIP, but not of CGRP, and cranial autonomic parasympathetic symptoms: A study in chronic migraine patients [Pubmed:27250233]
Headache No Change in Interictal PACAP Levels in Peripheral Blood in Women With Chronic Migraine [pubmed:27634731]
Sci Rep. Using the "target constituent removal combined with bioactivity assay" strategy to investigate the optimum arecoline content in charred areca nut. [pubmed:28054652]
BMC Complementary and Alternative Medicine [6]-Gingerol, from Zingiber officinale, potentiates GLP-1 mediated glucose-stimulated insulin secretion pathway in pancreatic β-cells and increases RAB8/RAB10-regulated membrane presentation of GLUT4 transporters in skeletal muscle to improve hyperglycemia in Leprdb/db type 2 diabetic mice [10.1186/s12906-017-1903-0]
Applied Biological Chemistry Positive enhancement of Lactobacillus fermentum HY01 on intestinal movements of mice having constipation [10.1007/s13765-017-0327-3]
Journal of Periodontology Salivary neuropeptides, stress, and periodontitis [Pubmed:28914594]
Evidence-Based?Complementary and Alternative Medicine Home-Based Transcutaneous Neuromodulation Improved Constipation via Modulating Gastrointestinal Hormones and Bile Acids [Pubmed:29853946]
Revista do Instituto de Medicina Tropical de S?o Paulo Vasoactive intestinal peptide degradation might influence Interleukin-17 expression in cardiac chagasic patients [Pubmed: 30365640]
Journal of Veterinary Science Serum Levels of Neuropeptides in Cows with Left Abomasal Displacement [Pubmed: 30562932]
Science?of?Food?and?Agriculture Protective Effect of Silkworm Pupa Oil on Hydrochloric Acid/Ethanol‐Induced Gastric Ulcer [Pubmed: 30479041]
MOLECULAR BIOLOGY REPORTS Serum CGRP, VIP, and PACAP usefulness in migraine: a case–control study in chronic migraine patients in real clinical practice [Pubmed: 32951099]
Journal of Cystic Fibrosis Disrupted local innervation results in less VIP expression in CF mice tissues [Pubmed: 32600901]
Biomedical Journal of Scientific & Technical Research Anti-Inflammatory and Pro-Inflammatory Neuropeptides in Dilative Cardiomyopathy are there Relations to Psychophysical Stress? []
Journal of tranditional chinese medical sciences Ganjiang granule regulates cecal microflora and serum biochemical components in a rat model of constipation-predominant irritable bowel syndrome []
J Cyst Fibros VIP reduction in the pancreas of F508del homozygous CF mice and early signs of Cystic Fibrosis Related Diabetes (CFRD) [34034984]
Neurol Sci Significance of serum VIP and PACAP in multiple sclerosis: an exploratory case–control study [34698942]
Endocr J A clinical analysis on functioning pancreatic neuroendocrine tumors (focusing on VIPomas): a single-center experience [Pubmed:35644576]
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